多头带绦虫dUTPase基因的原核表达、组织定位及酶学活性分析

doi:10.11843/j.issn.0366-6964.2017.12.017

Abstract

Abstract:

The aim of this study was to characterize the dUTPase gene of Taenia multiceps and analyze its enzymatic activity. The recombinant Tm-dUTPase was expressed in prokaryotic system and the enzymatic activity was measured; Western blotting analysis and immunofluorescence localization were also performed. Sequence analysis showed that the Tm-dUTPase gene contains a 447 bp open reading frame and the mature polypeptide consists of 148 amino acid residues, with a predicted molecular weight of 16.39 kDa and PI of 6.263; the deduced amino acid of Tm-dUTPase shares 97% and 91% similarity with dUTPase from Cysticercus cellulosae and Taenia pisiformis respectively. The recombinant Tm-dUTPase was expressed as a soluble protein, and the purified protein has a concentration of 0.907 mg·mL^-1. The Western blotting showed that the recombinant Tm-dUTPase could be recognized by the serum of goat naturally infected with Coenurus cerebralis, indicating a relatively high immunoreactivity. The immunolocalization showed that the native Tm-dUTPase mainly distributed to the parenchymatous zone of the adult parasite, and widely distributed to the protoscolexes and out layer of the cystic wall of Coenurus cerebralis. The enzyme activity of Tm-dUTPase was 986.29 U·mg^-1, and the activity could be inhibited by EDTA and enhanced by Mg²⁺, Zn²⁺ and Cu²⁺. These results would greatly benefit the further study of the biological function of Tm-dUTPase.

CLC Number:

S852.734

HUANG Xing, XU Jing, WANG Yu, YANG Ying-dong, WAN Jie, GUO Cheng, GU Xiao-bin, XIE Yue, YANG Guang-you. Prokaryotic Expression, Tissue Localization and Enzymatic Activity Analysis of Taenia multiceps dUTPase Gene[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(12): 2374-2382.

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Prokaryotic Expression, Tissue Localization and Enzymatic Activity Analysis of Taenia multiceps dUTPase Gene

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